anti gfp  (Novus Biologicals)

Journal: bioRxiv

Article Title: Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues

doi: 10.64898/2026.01.23.701393

Figure Lengend Snippet: a . Schematic of the tagged ENaCγ subunit produced from the ENaCγ-VF allele, which includes a C-terminal mVenus, 3C protease site, and 3xFLAG tag. b . Representative immunofluorescence image showing co-localization of mVenus and ENaCγ in kidney tissue. mVenus signal was detected using a FITC-conjugated anti-GFP primary antibody (goat, Rockland, 1:100) and Alexa Fluor 488 donkey anti-goat secondary antibody. Endogenous ENaCγ was detected using a rabbit anti-ENaCγ primary antibody (StressMarq, 1:500) targeting the C-terminus, and Alexa Fluor 647 donkey anti-rabbit secondary antibody. DAPI is 4’,6-diamidino-2-phenylindole . c . Close-up view highlighting localization of tagged ENaCγ.

Article Snippet: Representative immunofluorescence image showing co-localization of mVenus, ENaCα, and pNCC in kidney tissue. mVenus signal was detected using a FITC-conjugated anti-GFP primary antibody (goat, Rockland, 1:100) and Alexa Fluor 488 donkey anti-goat secondary antibody.

Techniques: Produced, Immunofluorescence

Journal: bioRxiv

Article Title: Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues

doi: 10.64898/2026.01.23.701393

Figure Lengend Snippet: a . Schematic workflow illustrating the experimental pipeline. Tissues (lung, kidney, colon) were harvested from ENaCγ-VF mice, homogenized in TRIS-based buffer, and followed by solubilization. Cleared supernatants were injected onto a Superose 6 Increase column for FSEC analysis, and fractions corresponding to high-molecular-weight complexes were subsequently analyzed by SiMPull. b . Representative FSEC traces showing mVenus fluorescence from lung, kidney, and colon lysates. Fluorescent peaks indicate the presence of γ-containing ENaC complexes in each tissue. c . Representative TIRF images of SiMPull experiments from lung lysates. mVenus-tagged ENaC complexes were captured using a biotinylated anti-GFP/mVenus nanobody. A background control image was collected from a flow chamber lacking the nanobody to demonstrate specificity. d . Quantification of SiMPull signal from lung, kidney, and colon tissues. Each condition represents data pooled from 40 TIRF images per sample, analyzed using the ComDet plugin in ImageJ. Statistical analysis was performed in GraphPad Prism.

Article Snippet: Representative immunofluorescence image showing co-localization of mVenus, ENaCα, and pNCC in kidney tissue. mVenus signal was detected using a FITC-conjugated anti-GFP primary antibody (goat, Rockland, 1:100) and Alexa Fluor 488 donkey anti-goat secondary antibody.

Techniques: Injection, High Molecular Weight, Fluorescence, Control

Journal: Journal of Virology

Article Title: Design of antibody structure-guided epitope vaccines in silico to induce potent immune responses against emerging viruses

doi: 10.1128/jvi.00689-25

Figure Lengend Snippet: Analysis of protein interactions of epitope peptides with scFv1. ( A ) Analysis of protein interaction between P30 peptide and scFv1. Plasmids pEGFP-P30 and pCMV-Flag-scFv1 were constructed and co-transfected into HEK293T cells. After 48 h, the proteins were collected and incubated with magnetic beads treated with mouse anti-GFP mAb. The samples were eluted with SDS-PAGE loading buffer and verified by Western blot. Plasmids pEGFP and pCMV empty vector were used as controls. ( B ) Analysis of protein interaction between S390F peptide and scFv1. ( C ) Analysis of protein interaction between G394K peptide and scFv1. ( D ) The self-activation verification of the bait plasmid PGBKT-scFv1 on SD-deficient medium. ( E ) Negative and positive controls in yeast two-hybrid assays. The pGADT-T plasmid was co-transformed with pGBKT-lam and pGBKT-53 plasmids in Y 2 Hgold competent cells as negative and positive controls, respectively. ( F ) The protein interactions between scFv1 and the P30, S390F, and G394K peptides were analyzed by yeast two-hybrid assay. After dilution, the bait plasmid pGBKT-scFv1 was co-transformed with the pGADT-P30, S390F, and G394K plasmids in Y 2 Hgold competent cells and cultured in SD-Leu-Trp and SD-Leu-Trp-His-Ade deficient medium for 48–96 h. ( G ) After a 10-fold dilution, the number of plaques on SD-LTHA deficient medium of competent cells co-transformed with pGADT-P30, S390F, and G394K with pGBKT-scFv1 plasmids was counted. * P < 0.05. ** P < 0.01. ( H ) After 100-fold dilution, the number of plaques on SD-LTHA deficient medium. * P < 0.05. ** P < 0.01. ( I ) After 1,000-fold dilution, the number of plaques on SD-LTHA deficient medium. * P < 0.05. ** P < 0.01. ( J ) The co-localization of P30, S390F, and G394K peptides with scFv1 was observed using laser confocal microscopy. To achieve this, pEGFP-P30, S390F, and G394K were co-transfected with pCMV-DsRed-scFv1 into HEK293T cells and cultured for 48 h. The fluorescence expression of the proteins was observed and analyzed under a 63× confocal microscope after DAPI staining with an anti-fluorescence quencher.

Article Snippet: In brief, TBS-washed protein A + G magnetic beads were incubated with GFP-tagged IgG2a mAb (1:3,000, Proteintech) for 1 h at room temperature.

Techniques: Construct, Transfection, Incubation, Magnetic Beads, SDS Page, Western Blot, Plasmid Preparation, Activation Assay, Transformation Assay, Y2H Assay, Cell Culture, Confocal Microscopy, Fluorescence, Expressing, Microscopy, Staining